Fig 8. Pharmacological blockage of endogenous IFN response increases the infection of HIE by HAstV from all clades.

A) HAstV1 (MOI = 1) infection of selected HIE derived from different human intestinal segments was carried out in the presence of trypsin and, when indicated, cells were further treated with 40 μM 2’-C-methylcytidine (2CMC) after adsorption. Viral genome copies were determined at 0 and 3 dpi by RT-qPCR. B) HAstV1 (MOI = 1) infection of selected HIE derived from different human intestinal segments was carried out in the presence of trypsin and pre-treated for 12 hrs with or without ruxotinilib (5 μM). Viral genome copies were determined at 0 and 3 dpi by RT-qPCR. C) Timecourse of MLB1 (MOI = 1) infection of C143 HIE in the presence or absence of ruxotinilib (5 μM) treatment. HIE was treated with ruxolitinib (5 μM) for 12 hrs before astrovirus infection. Virus genome copies were determined by RT-qPCR from extracted RNA at days 0, 1, 2 and 3 post infection. D) MLB1 (MOI = 1) infection of selected HIE pre-treated for 12 hrs with or without ruxotinilib (5 μM) was measured at 0 and 3 dpi. E) Timecourse of stool-derived HAstV (MOI = 1) infection of C68 HIE was carried out as described for HAstV-1. F) Infection of indicated HIE with stool-derived HAstV (MOI = 1) for 0 vs. 3 dpi as before. G-H) Effect of ruxotinilib (5 μM) treatment on G) IFN-β and H) ISG15 transcript expression in C68 HIE infected with stool-derived HAstV. Data are from ≥ 3 experiments; error = mean ± SD. Abbreviations: D = duodenum, J = jejunum, I = ileum, C = colon, IFN = interferon. The numbers indicate patient identifiers.